title: Mutation detection and typing of polymorphic loci through double-strand conformation analysis.
abstract: Variations, such as nucleotide substitutions, deletions and insertions, within genes can affect the function of the gene product and in some cases be deleterious. Screening for known allelic variation is important for determining disease and gene associations. Techniques which target specific mutations such as restriction enzyme polymorphism and oligonucleotide probe or PCR primer reactivity are useful for the detection of specific mutations, but these techniques are not generally effective for the identification of new mutations. Approaches for measuring changes in DNA conformation have been developed, based on the principle that DNA fragments which differ in nucleotide composition exhibit different mobilities after separation by polyacrylamide gel electrophoresis (PAGE). Here we describe a conformation-based mutation detection system, double-strand conformation analysis (DSCA), which provides a simple means to detect genetic variants and to type complex polymorphic loci. We demonstrate the application of DSCA to detect genetic polymorphisms such as a single-nucleotide difference within DNA fragments of up to 979 base pairs in length. We present the application of DSCA in detecting four different mutations in the cystic fibrosis gene (CFTR) and 131 different alleles encoded by HLA class I genes.
authors.affiliation: Anthony Nolan Research Institute, London, UK.
authors.forename: J R
authors.initials: JR
authors.lastname: Argüello
authors.forename: A M
authors.initials: AM
authors.lastname: Little
authors.forename: A L
authors.initials: AL
authors.lastname: Pay
authors.forename: D
authors.initials: D
authors.lastname: Gallardo
authors.forename: I
authors.initials: I
authors.lastname: Rojas
authors.forename: S G
authors.initials: SG
authors.lastname: Marsh
authors.forename: J M
authors.initials: JM
authors.lastname: Goldman
authors.forename: J A
authors.initials: JA
authors.lastname: Madrigal
mesh_terms: D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D006580:Genetic Carrier Screening; D015234:HLA-A Antigens / Q000235:genetics*; D015395:Histocompatibility Antigens Class I; D006801:Humans; D009154:Mutation; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic
chemical_list: D015234:HLA-A Antigens; D015395:Histocompatibility Antigens Class I; D004247:DNA